A fusion protein of glucagons-like peptide-1 and human serum albumin (GLP-1/HSA) was expressed and secreted into the fermentation broth with recombinant Pichia pastoris. The productivity of expressed GLP-1/HSA could reach 63.6mg/L in 10L fermentor. After concentrated with hollow-fiber ultrafiltration membrane, GLP-1/HSA was purified from fermentation broth by hydrophobic chromatography, negative ion exchange chromatography and gel filtration chromatography in turn. The HPLC analysis showed that the purified GLP-1/HSA had an overall purity of 95.8%. Furthermore, the analysis of in vivo activity indicated that GLP-1/HSA had the bioactivity of native GLP-1, and could significantly reduce blood glucose level 4h after intraperitoneal administration. It was concluded that a great deal of GLP-1/HSA with higher purity could be harvested by Pichia pastoris expression system and the established purification methods. Preliminary studies show a new potential for developing the long-acting GLP-1 analogs for clinical applications.

BACKGROUND:Pathophysiological mechanisms after spinal cord injury are very complex, so there is no compressive and in-depth understanding on it.
OBJECTIVE:To study the effect of dura mater spinalis integrity on cytokine levels in the cerebrospinal fluid of animal models of spinal cord injury.
METHODS:The white rabbit models of spinal cord injury were established using clamp compression method, and then the models were randomly divided into four groups:no dura mater spinalis defect group, dura mater spinalis defect group, dura mater spinalis defect composite with membrane repairing group and dura mater spinalis defect composite with autologous fascia repair group. Enzyme-linked immunosorbent assay was
performed to detect the changes of levels of cytokines (interleukin-6, interleukin-10 and tumor necrosis factorα) in the cerebrospinal fluid at 30 minutes, 1, 3, 6, 12 and 36 hours after surgery.
RESULTS AND CONCLUSION:The levels of interleukin-6, interleukin-10 and tumor necrosis factorαin the
cerebrospinal fluid of the dura mater spinalis defect group, dura mater spinalis defect composite with membrane repairing group and dura mater spinalis defect composite with autologous fascia repair group were significantly lower than those of the no dura mater spinalis defect group at 6 hours after surgery (P<0.05). There were no significant differences in the levels of interleukin-6, interleukin-10 and tumor necrosis factorαat other time points between groups (P>0.05). The results indicate that maintaining the integrity of dura mater spinalis of the spinal cord injury model can affect the levels of interleukin-6, interleukin-10 and tumor necrosis factorαin the
cerebrospinal fluid, thus inhibiting the inflammatory response.

Objective To find a proper number of implanted fiducials in order to reconcile both more accurate image tacking and less pain for the patients.Methods The phantom was made of two parts of different materials.The inner part was a ball-cube phantom developed by Accuray Corporation, which could be regarded as a rigid body, with 5 fiducials inside using as the reference to align the whole phantom in the experiment.The outer part was made of additional silicone rubber with two components to simulate soft tissue as a non-rigid body, which was implanted with 8 fiducials inside.All combinations of different number of fiducials were tracked to aquire the target location information, which were then compared with the reference position that was set by the inner 5 fiducials to obtain the deviations of the translation and the rotation parameters.Thus the impact of the number of fiducials on image target localization accuracy could be analyzed easily.Results When 4 fiducials are used for tracking, the decline of translation and rotation errors for every fiducial is largest (0.086 mm and 0.033°).The translation error was decreased by 0.343 mm from 1 to 4 fiducials, while by only 0.077 mm from 4 to 8 fiducials;the same decline was observed for rotation errors (0.131° for 3 to 4 fiducials , and 0.09° for 4 to 8 fiducials , respectively) .Conclusions When 4 fiducials are used for tracking, the decline of errors for every fiducial is maximum.When one more fiducial is adding, the image target localization accuracy can be enhanced obviously from 1 to 4 fiducials, but not as obvious for more than 4 fiducials.

Objective To explore the appropriate treatment according to the grading system of adolescent idiopathic cervical kyphosis.Methods A retrospective study was performed in 115 adolescent patients with idiopathic cervical kyphosis.The patients were divided into 4 groups according to the magnitude of kyphosis.The initial Cobb angle of 4 groups were 12.7°±1.4° 25.4°±4.8°,47.2°±4.4° and 62.6°±5.7° respectively.The patients in group I were treated with the collar support for 4-8 weeks.The patients in group Ⅱ were treated with skull traction (3-5 kg) and then fixed by cranio-cervical-thoracic plaster.According to the angles between the tangents of posterior vertebral body at each level on lateral cervical radiograph in extension,the anterior fusion levels of the group Ⅲ and angles and range of osteotomy in the group Ⅳ were decided.In group Ⅳ,the patients were treated by two steps.The anterior release and posterior osteotomy were performed firstly.Then skull traction (1/10 body weight) was maintained in order to correct the deformity for 7-10 days,fusion and anterior fusion with autologous bone graft and internal fixation was completed.Results Post-operative radiograph showed that Cobb angle were -5.5°±2.0°,-8.2°±6.1°,-4.5°±6.6° and -2.9°±7.9° in Ⅰ-Ⅳ group after treatment.The deformed appearance of the patients improved significantly.A improvement neck pain and neurologic function were found in all patients.Post-operative MRI showed that physiological curve of the cervical spine was restored,and the cerebrospinal fluid line was clear in the previous kyphosis area.Conclusion Adolescent idiopathic cervical kyphosis has specific characteristics.Surgical strategy is determined by the severity of deformity.

Objective:To explore the propaganda methods and propagation path of advertising media cosmetology publicity in accordance with public demand and ethical principles,through understanding college students' acceptance level for advertising media cosmetology publicity and its influence on their choice forcosmetic ways.Methods:With the method of stratified cluster sampling,an anonymous questionnaire survey was conducted among 210 college students in a university in Shaanxi province.Results:The Internet has become the main way for college students to acquire cosmetology knowledge.A total of 44.7 ％ of the students thought most of the media cosmetology publicity was unbelievable,46.5％ of the students thought that advertising media publicity mainly considered the interests of the beauty institution,72.9％ of the students would not consider the beauty institution the media recommended,62.4％ of the students would choose regular public hospitals,56.3％ of the students would give up the idea of cosmetology due to the news of cosmetic failure,42.2％ of the students could accept minimally invasive plastic surgery,and 68.5 ％ of the students expected to acquire the cosmetology knowledge through expert consultation.Concltusion:At current,the credibility of media cosmetology publicity is not optimistic enough.It should further strengthen the supervision for media cosmetology publicity.Cosmetology advertising media should construct the publicity mode based on ethics,law,governance,occupation,public welfare and social responsibility.

Objective To explore the efficacy of mycophenolate mofetil (MMF), which is a kind of immuno-suppressant drugs, on the treatment of Connective tissue diseases-pulmonary arterial hypertension (CTD-PAH). Methods Medical charts of eleven cases of hospitalized patients who were diagnosed as CTD-PAH and treated by MMF in Beijing Chaoyang Hospital affiliated to the Capital Medical University, from January 2014 to June 2016 were collected and analyzed. Results In the 11 cases of CTD-PAH, the systemic lupus erythematosus (SLE) related pulmonary hypertension (SLE-PAH) were 7, while the systemic sclero-derma associated pulmonary hypertension (SSc-PAH) were 2, and rheumatoid arthritis related pulmonary hy-pertension (RA-PAH) was 1, and the mixed connective tissue disease related pulmonary hypertension (MCTD-PAH) was 1. All patients were women, and the average age was (40 ±14) years, and the average duration of PAH was (34 ±35) months. The combination therapy of corticosteriods and MMF was applied to 7 cases, meanwhile the therapy of corticosteriods, MMF and bosentan was used in 1 case, corticosteriods, MMF and sildenafil was prescribed for 3 cases, and symptoms of the patients alleviated. Except for one case having been followed up for 7 months and one for 6 months, 9 patients completed the 1-year follow-up, and the survival rate was 100%(9/9). Notably, one patient, who had been alleviated for 111 months with therapy of corticosteriods and MMF, adopted the combination therapy of corticosteriods, MMF and bosentan for aggravated chest distress, and became stable eventually. Conclusion MMF may have therapeutic effects on inducing and even maintaining the stabilization of CTD-PAH.

Objective To screen mutations in genes including ASXL1, TET2, IDH1, IDH2, SETBP1, MPL515, JAK2 exon 12 and JAK2V617 in 135 polycythemia vera (PV) patients. To assess progreasson and genomics characteristics post polycythemic myelofibrosis. Methods DNA sequencing of ASXL1(Exon12),TET2 (Exons 3-11),IDH1(Exon4),IDH2(Ex-on4),SEPBP1(Exon4),JAK2 exon 12 and MPL515 (Exon 10) genes were carried out using Sanger method. JAK2V617 muta-tion was detected by allele-specific PCR in patients with PV. In the mean time, the mutation load of JAK2V617F allele (V617F%) was evaluated by real-time PCR using Tagman MGB probe. Then, the significant of gene mutations and clinical outcomes of post-PV Myelofibrosis(PPMF)was analyzed. To study risk factors of PPMF, logistic regression were employed. Results ASXL1, TET2, IDH1, IDH2 were mutated in 7.69%(8/104), 5.26%(1/19) , 0.08%(1/120) and 0.08%(1/121) of all PV patient respectively. JAK2 was mutated in 82.22%(111/135) of PV patients with mutation rate of exon12 of 2.96%(4/135) and there were no mutation of MPL515 and SETBP1 in PV patients. ASXL1 mutation was found in 31.82%(7/22) PPMF patients. Spearman analysis showed that ASXL1 is correlated with JAK2V617F (V617F%) (rs=0.298,P=0.002). The hemo-globin was lower in patients with ASXL1 mutation than patient without mutation (wild type). Leukocyte count, V617F%>50%rate, thrombosis and PPMF were higher in patients with ASXL1 mutation than that of ASXL1 wild type(P＜0.05). ASXL1 mu-tation, V617F%>50% rate and splenomegaly were all risk factors of PPMF. Conclusion ASXL1 mutation is the risk-fac-tor of PPMF and may promote V617F%by some mechanism.

Objective To compare the degree of cell injury induced by 3D protein (SDLY11 and SDLY107) of enterovirus 71 (EV71) strains.Methods EV71 strains SDLY11 and SDLY107 were respectively isolated from children with mild and severe hand foot mouth disease.The target genes 11-3D-Flag and 107-3D-Flag were amplified by reverse transcription-polymerase chain reation (RT-PCR) and inserted into the eukaryotic expression vector pcDNA3.1.The recombinant plasmids 11-3D-Flag-pcDNA3.1 and 107-3D-Flag-pcDNA3.1 were transformed into Escherichia.coli DH5α, respectively, and were identified by enzyme digestion and sequencing.The recombinant plasmids were transfected into rhabdomyosarcoma (RD) cells, respectively.Expression of 3D protein was detected by indirect immunofluorescence assay and western blot.Cell injury induced by 3D protein was detected with lactate dehydrogenase (LDH) test, cell proliferation was detected by 3-(4,5-dimethylthiazolyl-2)-2,5-diphenylthiazolium bromide (MTT) test, and cell apoptosis was detected with Annexin-V and PI.Multiple comparisons among groups were analyzed using LSD-t test if multiple sets of variables were consistent with homogeneity of variance.If not, Dunnett T3 test was used.Results The 1 400 bp fragments were amplified by reverse tramscription (RT)-polymerase chain reaction (PCR), and the recombinant plasmids were digested by enzyme and the 1 400 bp and 5 400 bp fragments were obtained and identified.Gene sequencing showed that the sequences were consistent with the target genes.The specific fluorescence was observed by indirect immunofluorescence assay, and the western blot showed that the molecular weight of the target protein was 55×103.The LDH test showed that the A490 of SDLY11 3D protein transfection group (0.790±0.048) was higher than that of SDLY107 3D protein transfection group (0.641±0.018).The difference was statistically significant (t=5.14, P<0.05).The cell membrane damage caused by SDLY11 3D protein was more severe than SDLY107 3D protein.The MTT test showed that the A570 of SDLY11 3D protein transfection group (1.028±0.020) was lower than that of SDLY107 3D protein transfection group (1.081±0.002), and the difference was statistically significant (t=3.31, P<0.05).The effect on cell proliferation activity of SDLY11 3D protein was greater than SDLY107 3D protein.The results of Annexin-V/PI showed that the percentage of apoptotic cells of SDLY11 3D protein transfection group and SDLY107 3D protein transfection group were (1.471±0.246)% and (1.465±0.237)%, respectively, and the difference was not statistically significant (t=0.04, P=0.973).Conclusions Compared with the SDLY11 3D protein, SDLY107 3D protein induces slighter cell injury, has weaker effect on cell proliferation activity, and is more favorable for virus replication in cells.

Lu Dangshen, a traditional authentic medicinal material of Codonopsis Radix is mainly produced in Shangdang (Changzhi) area of Shanxi Province. Baitiao Dangshen is mainly produced in Gansu Province. Codonopsis Radix contains many kinds of components such as phenylpropanoids, polyalkynes, alkaloids, terpenes, fatty acids, flavonoids, and so on. At present, the effect of producing areas on its chemical compositions has not been systematically studied. This study analyzed the differences of metabolites among Codonopsis pilosula from different producing areas by UPLC-HRMS. PCA, OPLS-DA coupled with Thermo mzcloud online and local databases were used to compare the overall differences of metabolites among Codonopsis pilosula from different producing areas, and the chemical constituents were identified to further screen and find out the different metabolites and analyze the metabolic pathways by information retrieval in HMDB, PubChem, Chemspider and KEGG databases. The results showed that 72 differential metabolites were identified in this study. There were 15 kinds of up-regulated and 57 kinds of down-regulated metabolites of Lu Dangshen compared with Baitiao Dangshen. The top 30 metabolic pathways were analyzed by KEGG enrichment, and the most important metabolic pathways were phenylpropanoid biosynthesis, which was demonstrated that phenylpropanoid biosynthesis pathway and related intermediate metabolites could be used as the characteristics of distinguishing Lu Dangshen from different habitats of Codonopsis pilosula. The present study provided a basis for analyzing the influence of producing areas on the chemical components of Codonopsis pilosula and reasonably evaluating the quality of Codonopsis Radix, and also provided a new idea for expounding the authenticity of Lu Dangshen.

BACKGROUNDHepatitis C virus (HCV) infection may induce autoimmune response and autoantibodies can be detected in chronic hepatitis C (CHC) patients. However, the reported positive rate of autoantibodies in CHC patients in China varies considerably. In this study, we investigated the prevalence of antinuclear antibodies (ANA) and anti-liver-kidney-microsome type 1 autoantibodies (anti-LKM-1) in a large cohort of CHC patients, and analyzed the factors related to the presence of the autoantibodies.

METHODSA total of 360 CHC patients were enrolled in this study. Serum ANA and anti-LKM-1 were detected by indirect immunofluorescence and enzyme-linked immunosorbent assay, respectively. Clinical analysis was performed to disclose the related factors to autoantibody production.

RESULTSThe prevalence of ANA and anti-LKM-1 in CHC patients was 12.5% (45/360) and 2.5% (9/360), respectively. Women had a higher prevalence than men (18.9% vs 11.4%, P = 0.046). Patients with positive autoantibodies had lower HCV RNA levels (1.2 x 10(7) copies/L vs 7.2 x 10(7) copies/L, P < 0.05). Positive ANA was associated with higher serum globulin (P < 0.05). Stratified analysis showed that there were no significant differences in age, HCV genotype, disease course, clinical stage, prevalence of cirrhosis and interferon therapy between autoantibody-positive and -negative subgroups.

CONCLUSIONAutoantibodies can be induced in the course of CHC, and some CHC patients can even develop autoimmune hepatitis.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antibodies, Antinuclear ; blood ; Autoantibodies ; blood ; Child ; Child, Preschool ; China ; Enzyme-Linked Immunosorbent Assay ; Female ; Fluorescent Antibody Technique, Indirect ; Hepatitis C, Chronic ; blood ; immunology ; Humans ; Male ; Middle Aged ; Prevalence ; Young Adult